Eosin, also known as tetrabromofluorescein, is a xanthene dye among synthetic dyes, appearing as a peach red or pink powder with the molecular formula C₂₀H₆O₅Br₄Na₂ and a molecular weight of 691.86. Clinically, sperm viability can be assessed by detecting the integrity of the sperm membrane. Aladdin Sperm Vital Staining Solution (Eosin Method) enables routine viability testing of all sperm samples. It identifies sperm with intact cell membranes via the dye exclusion test, thereby determining the percentage of viable sperm. Composed of eosin, deionized water and other components, this staining solution is based on the principle of dye exclusion: damaged cell membranes, such as those of non-viable (dead) cells, allow impermeant dyes to penetrate and stain the intracellular space, whereas the cell membranes of viable cells can resist the entry of dyes, resulting in the dye exclusion phenomenon with no staining observed. This reagent is for research use only and not intended for clinical diagnosis or any other applications.
Operating Procedures (For Reference Only):
1. Take a clean glass slide, add 1 drop each of fresh semen and Sperm Vital Staining Solution (Eosin Method), and mix well.
2. Cover with a coverslip, let stand for 30–60 seconds, and observe immediately under a high-power microscope (400× recommended).
3. Count 200 sperm cells and calculate the percentage of unstained sperm (viable sperm) among the total 200.
Staining Results
Viable sperm Unstained (white)
Non-viable sperm Red
Precautions:
1. Sperm viability testing should be performed immediately after semen liquefaction, preferably within 30 minutes and no more than 1 hour under any circumstances. Delayed testing may cause sperm inactivation due to dehydration or temperature changes, leading to inaccurate staining results.
2. Use the reagent as soon as possible after opening to avoid compromising subsequent experimental outcomes.
3. For your safety and health, wear a lab coat and disposable gloves during operation.
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